As part of a Ministry of Science, Technology, and Innovation initiative, the Brazilian Societies of Botany, Microbiology, Virology, and Zoology conducted a comprehensive evaluation of biological collections in Brazil. This assessment aimed to gather insights into the current state of these collections, with the goal of providing support for future public policies, including financial subsidies and prioritization policies. In this context, we present the findings related to microbiological collections, essential to ex situ biodiversity conservation and crucial in supporting research, development, and innovation. A survey was distributed to public and private institutions across Brazil, yielding responses from 168 microbiological collections representing 79 different entities. Notably, 73 of these collections are affiliated with public research institutions and universities, underscoring the State’s pivotal role in preserving and safeguarding Brazilian microbial diversity. The primary taxonomic groups encompass bacteria (found in 70.24% of collections) and fungi (comprising 52.98% of collections), sourced from diverse Brazilian ecosystems and biomes, including those that contain several type strains. Furthermore, the collections preserve microorganisms harboring biotechnological potential applicable to environmental protection, public health, industry, and agribusiness. Despite these promising economic and biotechnological prospects, our meticulous data analysis has revealed significant limitations and vulnerabilities, especially regarding physical infrastructure and human resources, emphasizing the urgent need for interventions to guarantee their sustainability.